Esterase-activated two-fluorophore system for ratiometric sensing of biological zinc(II)

被引:96
作者
Woodroofe, CC [1 ]
Won, AC [1 ]
Lippard, SJ [1 ]
机构
[1] MIT, Dept Chem, Cambridge, MA 02139 USA
关键词
D O I
10.1021/ic048789s
中图分类号
O61 [无机化学];
学科分类号
070301 ; 081704 ;
摘要
Intracellular ester hydrolysis by cytosolic esterases is a common strategy used to trap fluorescent sensors within the cell. We have prepared analogues of Zinpyr-1 (ZP1), an intensity-based fluorescent sensor for Zn2+, that are linked via an amido-ester or diester moiety to a calibrating fluorophore, coumarin 343. These compounds, designated Coumazin-1 and -2, are nonpolar and are quenched by intramolecular interactions between the two fluorophores. Esterase-catalyzed hydrolysis generates a Zn2+ -sensitive ZP1-like fluorophore and a Zn2+ insensitive coumarin as a calibrating fluorophore. Upon excitation of the fluorophores, coumarin 343 emission relays information concerning sensor concentration whereas ZP1 emission indicates the relative concentration of Zn2+-bound sensor. This approach enables intracellular monitoring of total sensor concentration and provides a ratiometric system for sensing biological zinc ion.
引用
收藏
页码:3112 / 3120
页数:9
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